【摘要】：樹突狀細胞(dendritic cell, DC)是已知體內功能最強,唯一能活化初始型T 細胞的專職抗原遞呈細胞(antigen presenting cell, APC),其數量、成熟狀態(tài)可決定機體免疫狀態(tài),故對其深入的研究具有重要理論和實際意義。DC 具有體外可操作性,因此獲取一定量有功能的DC 是深入研究其生物特性的關鍵。 本研究目的為在體外建立一個穩(wěn)定的從人外周血單核細胞誘導未成熟及成熟DC 的培養(yǎng)方法,經流式細胞術檢測細胞表面分子的表達情況并應用MTT 法對培養(yǎng)細胞的生物學功能進行測定,從而確保獲得具有一定形態(tài)和功能特征的DC,為進一步探討DC 在免疫耐受及抗腫瘤方面作用奠定基礎。 研究結果:本研究建立了誘導外周血單核細胞獲得不同發(fā)育階段DC的方法,獲得較大量,純度較高的CD83+DC。與以往報道有所不同,經流式細胞術檢測高表達CD83分子的DC仍有CD14分子的表達。經過再次混合淋巴細胞反應發(fā)現具有中度表達共刺激分子的未成熟DC在體外可以誘導同一來源的T細胞免疫低應答。 通過本研究我們建立了穩(wěn)定的體外DC 培養(yǎng)體系,為移植免疫耐受研究和抗腫瘤研究深入發(fā)展提供了理想的基因轉移靶細胞。
[Abstract]:Dendritic cell (dendritic cell, DC) is known to have the strongest function in vivo. The only specialized antigen-presenting cell (antigen presenting cell, APC),) that activates the initial T cell is the number of (antigen presenting cell, APC), and the mature state determines the immune state of the body. Therefore, the in-depth study of DC has important theoretical and practical significance. DC can be operated in vitro, so obtaining a certain amount of functional DC is the key to further study its biological characteristics. The aim of this study was to establish a stable culture method for inducing immature and mature DC from human peripheral blood monocytes in vitro. The expression of cell surface molecules was detected by flow cytometry and the biological function of cultured cells was determined by MTT assay. So as to ensure that DC, with certain morphological and functional characteristics lay a foundation for further study of the role of DC in immune tolerance and antitumor. Results: in this study, we established a method to induce peripheral blood monocytes to obtain DC at different developmental stages. A large number of CD83 DC. with high purity were obtained. Different from previous reports, DC with high expression of CD83 still expressed CD14 by flow cytometry. After remixed lymphocyte reaction it was found that immature DC with moderate expression of costimulatory molecules could induce low immune response of T cells from the same source in vitro. Through this study, we established a stable culture system of DC in vitro, which provides an ideal target cell for gene transfer for the further development of transplantation immune tolerance and antitumor research.
【學位授予單位】：吉林大學
【學位級別】：碩士
【學位授予年份】：2005
【分類號】：R392

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